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Complete genome sequence and amplification primer of Seneca valley virus SVV/CH/NM/2016

A whole-genome, Seneca technology, applied in the field of molecular biology, can solve the major risks and economic losses of infected pigs in the market

Active Publication Date: 2018-05-29
JINYUBAOLING BIO PHARMA CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

From November 2014 to April 2015, there were several SVV-infected pig herds in Brazil with severe clinical morbidity and death symptoms, causing serious economic losses
[0004] Since Seneca Valley Virus (SVV) infection clinically resembles Foot-and-Mouth Disease (FMD), there is a significant risk in bringing infected pigs to market

Method used

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  • Complete genome sequence and amplification primer of Seneca valley virus SVV/CH/NM/2016
  • Complete genome sequence and amplification primer of Seneca valley virus SVV/CH/NM/2016
  • Complete genome sequence and amplification primer of Seneca valley virus SVV/CH/NM/2016

Examples

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Embodiment 1

[0041] Example 1. Primer combination designed for one-step RT-PCR amplification of 7 nucleotide sequence fragments of the whole genome of Seneca Valley virus SVV / CH / NM / 2016

[0042] One of the objects of the present invention is to obtain the whole genome sequence of Seneca Valley virus SVV / CH / NM / 2016, so that the molecular genetic evolution trend and epidemic situation of Seneca Valley virus are more comprehensive and comprehensive at the level of the whole genome sequence. Learn systematically for further in-depth research.

[0043] According to the full genome nucleotide sequence of Seneca Valley virus reference strains in NCBI, such as SVA / CH / 01 / 2015 (GenBank: KT321458.1), SVA / CH / 02 / 2015 (GenBank: KX173339.1), SVA / CH / DL / 01 / 2016(GenBank:KX751944.1), SVA / CH / GXI09 / 2016(GenBank:KY038016.1), SVA / CH / LX / 01 / 2016(GenBank:KX751945.1), SVA / CH / ZW / 01 / 2016(GenBank:KX751946.1), SVA / HLJ / CHA / 2016(GenBank:KY419132.1), SVA / CH / FJ / 2017(GenBank:KY747510.1), SVA / CH / HN / 2017(GenBank:KY747511.1...

Embodiment 2

[0047] Example 2, obtaining the whole genome sequence of Seneca Valley virus SVV / CH / NM / 2016

[0048] Based on the primer combination obtained in Example 1, the present invention can obtain the whole genome sequence of Seneca Valley virus SVV / CH / NM / 2016, and the obtaining method comprises the following steps:

[0049] 1. Extract the total RNA of Seneca Valley virus SVV / CH / NM / 2016

[0050] Follow Axyprep TM Body Fluid Viral DNA / RNA Miniprep Kit (purchased from AXYGEN Company) instructions, extract the total RNA of Seneca Valley virus SVV / CH / NM / 2016 (preservation number: CGMCC 14885), the specific steps are as follows:

[0051] (1) Reagent preparation: Prepare isopropanol containing 1% (V / V) glacial acetic acid in advance; add specified volumes of absolute ethanol to reagent Buffer W1A and Buffer W2 respectively. That is, add 17mL absolute ethanol to 24mL Buffer W1A; add 56mL absolute ethanol to 24mL Buffer W2.

[0052] (2) Take 200 μL of the sample to be tested (the venom of...

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Abstract

The invention discloses a complete genome sequence and an amplification primer of a Seneca valley virus SVV / CH / NM / 2016. A one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method is firstly adopted for amplifying seven nucleotide sequence fragments (S1, S2, S3, S4, S5, S6, and S7) of an SVV / CH / NM / 2016 strain, then the seven nucleotide sequence fragments are cloned and sequenced, and then, the DNA sequences of the seven nucleotide sequence fragments are sequentially spliced, edited and corrected to finally obtain the complete genome sequence of the Seneca valley virus SVV / CH / NM / 2016. The obtained complete genome sequence of the Seneca valley virus SVV / CH / NM / 2016 is favorable for further research of the pathogenic mechanism of the Seneca valley virus, molecular epidemiology,reverse genetics and the like, thereby establishing important data support and theoretical basis for diagnostic reagent development, vaccine development and the like of the Seneca valley virus.

Description

technical field [0001] The invention belongs to the whole genome sequence of Seneca Valley virus in the field of molecular biology, in particular to the whole genome sequence of Seneca Valley virus SVV / CH / NM / 2016 and its amplification primers. Background technique [0002] Seneca Valley Virus (SVV) is also known as Seneca Valley Virus (SVV), Porcine Seneca Valley Virus (SVV), Senecavirus A (SVA). SVV is a single-stranded, non-segmented, and non-enveloped RNA virus, which is the only member of the Picornaviridae Senecavirus genus. The SVV virus particle is typical icosahedral symmetry, with a diameter of about 27nm and a molecular weight of about 30KD. The total length of the SVV genome is about 7.2kb, including two conserved non-coding regions 5'-UTR, 3'-UTR and an open reading frame (ORF), ending with a poly(A) at the 3' end. [0003] Seneca Valley virus (SVV) is an emerging virus that infects and kills piglets and sows. SVV infection can cause blisters and ulcerated wou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/41C12N15/11C12N15/10C07K14/085
CPCC12Q1/686C07K14/005C12N2770/32022
Inventor 陈君彦王秀明魏学峰刘国英关平原陈九连范秀丽张燕红王云凌张贵刚王艳杰张宸刘建奇武瑾贤谢雪岑杜宇荣
Owner JINYUBAOLING BIO PHARMA CO LTD
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